This invention relates to the identification of diarylsulfonylurea (DASU) binding proteins (DBPS) as novel therapeutic targets for suppressing the release of inflammatory mediators such as interleukin IL-1 and IL-1xcex2.
Inflammatory diseases such as rheumatoid arthritis are characterized by an excessive production of cytokines that promote and/or maintain the inflammatory state. Prominent among them are IL-1 (both the xcex1 and xcex2 forms), tumor necrosis factor alpha (TNFxcex1), and IL-18 (Dinarello, C. A. Blood 87:2095-2147 (1996); Aggarwal, B. B. and Natarajan, K. Eur Cytokine Netw. 7:93-124 (1996); Ushio, S. et al. J. Immunol. 156:4274-4279 (1996)). After release from producing cells, these cytokines bind to specific receptors on target cells to initiate cytokine signaling cascades. As a result of their importance in the disease process, therapeutic approaches aimed at regulating production and/or activity of these cytokines are desirable.
Most cytokines are secreted from cells via the constitutive secretory apparatus composed of the rough endoplasmic reticulum and Golgi apparatus, but IL-1 and IL-18 are exported by an untypical route (Rubartelli, A. et al. EMBO J. 9:1503-1510 (1990)). The need for a non-traditional export pathway is a consequence of the synthesis of IL-1 and IL-18 as polypeptides lacking signal sequences (Auron, P. E. et al. Proc. Natl. Acad. Sci. USA 81:7907-7911 (1984); March, C. J. et al. Nature 315:641-647 (1985); Ushio, S. et al. J. Immunol. 156:4274-4279 (1996)). This signal or leader sequence typically is found at the amino terminus of polypeptides which are destined to be released from the cell (von Heijne, G. J. Membrane Biol. 115:195-201 (1990)). A signal sequence serves as a molecular address to direct newly synthesized polypeptides into the endoplasmic reticulum. Because newly synthesized IL-1 (prolL-1) and IL-18 (prolL-18) lack this sequence, they accumulate within the cytoplasmic compartment of the producing cell. In addition to their co-localization, prolL-1xcex2 and prolL-18 also must be processed by the protease caspase I (Thomberry, N. A. et al. Nature 356:768-774 (1992); Ghayur, T. et al. Nature 360:619-623 (1997)); this cleavage generates biologically active, mature forms of the cytokines competent to bind to target cell receptors. ProlL-1xcex1 does not share this requirement for proteolytic activation (Moseley, B. et al. J. Biol. Chem. 262:2941-2944 (1987). Other polypeptides exported by similar non-traditional routes include: Mif-related protein (MRP) 8/14 (Rammes, A. et al. J. Biol. Chem. 272:9496-9502 (1997)), basic fibroblast growth factor (Abraham, J. A. Science 233: 545-548 (1986)), galectins (Cleves, A. E. et al. J. Cell Biol. 133:1017-1026 (1997)), a form of the IL-1 receptor antagonist (Haskill, S. et al. Proc. Natl. Acad. Sci. USA 88:3681-3685 (1991)), thioredoxin (Rubartelli, A. et al. J. Biol. Chem. 267:24161-24164 (1992)), and several virally-encoded polypeptides including VP22 and Tat (Ensoli, B. et al. J. Virol. 67:277-287 (1993)); Elliott, G. and O""Hare, P. Cell 88:223-233 (1997)).
Lipopolysaccharide (LPS)-treated monocytes and macrophages produce large quantities of prolL-1xcex2, but the release of mature cytokine is inefficient in the absence of a secondary stimulus (Hogquist, K. A. et al. J. Immunol. 147:2181-2186 (1991)); Perregaux, D. et al. J. Immunol. 149:1294-1303 (1992)). Both proteolytic maturation of prolL-1xcex2 and the release of mature cytokine are enhanced by treating LPS-activated cells with any of a number of different stimuli including: extracellular ATP, cytolytic T-cells, high concentrations of LPS, ionophore-like molecules, toxins, hypotonic stress, and mechanical stress (Hogquist, K. A. et al. Proc. Natl. Acad. Sci. USA 88:8485-8489 (1991); Perregaux, D. and Gabel, C. A. J. Biol. Chem. 269:15195-15203 (1994); Walev, I. et al. Eur. Mol. Biol. Org. J. 14:1607-1614 (1995); Bhakdi, S. et al. J. Clin. Invest. 85:1746-1753 (1990); Chin, J. and Kostura, M. J. J. Immunol. 151:5574-5585 (1993)). Importantly, stimulus-coupled cytokine posttranslational processing is sensitive to pharmacological intervention. Thus, a variety of non-selective anion transport inhibitors such as ethacrynic acid and tenidap can block stimulus-coupled posttranslational processing of prolL-1xcex2 (Laliberte, R. et al. J. Immunol. 153:2168-2179 (1994); Perregaux, et al. J. Immunol. 157:57-64 (1996); Perregaux et al. J. Immunol. 160:2469-2477 (1998)). These agents are effective inhibitors of IL-1 posttranslational processing independent of the nature of the activating stimulus. Moreover, their inhibitory effect is manifested as a complete suppression in the externalization of both IL-1xcex1 and IL-1xcex2.
A novel series of agents termed DASUs has been identified as potent inhibitors of stimulus-coupled posttranslational processing. These compounds are described and claimed in United States Provisional Application No. 1,086,939 filed Jan. 27, 1997, the entire disclosure of which is hereby incorporated by reference. Because IL-1 and IL-18 are important mediators of inflammation and inhibitors of their function provide therapeutic relief in animal models of disease (Cominelli, F. et al. J. Clin. Invest. 86:972-980 (1990); Akeson, A. L. et al. J. Biol. Chem. 271:30517-30523 (1996); Caron, J. P. et al. Arthritis Rheum. 39:1535-1544 (1996); Okamura, H. et al. Nature 378:88-91 (1995); Rothwell, N. J. Clin. Invest. 100:2648-2652 (1997)), it is anticipated that agents that disrupt the process of stimulus-coupled posttranslational processing will be useful for the treatment in man and animals of disorders that are sustained by inflammatory mediators. These include rheumatoid arthritis, osteoarthritis, asthma, inflammatory bowel disease, ulcerative colitis, neurodegeneration, atherosclerosis, and psoriasis.
This invention relates to the identification of protein and DNA sequences corresponding to two DASU binding proteins (DBPs) that mediate the cytokine inhibitory activity of these agents. DBPs may be used to screen for structurally unique drugs that disrupt stimulus-coupled posttranslational processing. Compounds that bind to the DBPs also may be used as therapeutics in the treatment of inflammatory disorders.
The invention further relates to a pharmaceutical composition of a drug that binds to the DBPs.
The following abbreviations are used throughout this patent:
In one embodiment, the present invention is directed to a method of screening for the ability of a compound to inhibit the production of an inflammatory cytokine comprising determining the ability of the compound to bind to a polypeptide coded for by the polynucleotide sequence of SEQUENCE ID NO: 1, or a polypeptide coded for by a polynucleotide sequence having 95% homology to SEQUENCE ID NO: 1.
Especially preferred is the method wherein the inflammatory cytokine is interleukin 1.
In another embodiment, the present invention is directed to method of treatment of a mammal having a disease condition characterized by an inflammatory component comprising administering to a mammal in need of such treatment a compound, or a pharmaceutically acceptable salt, ester, or prodrug of said compound, capable of binding to a polypeptide coded for by the polynucleotide sequence of SEQUENCE ID NO: 1, or a polypeptide coded for by a polynucleotide sequence having 95% homology to SEQUENCE ID NO: 1.
In yet another embodiment, the present invention is directed to a pharmaceutical composition comprising a pharmaceutical carrier and a pharmaceutically active amount of a compound, or a pharmaceutically acceptable salt, ester, or prodrug of said compound, said compound capable of binding to the polypeptide coded for by the polynucleotide sequence of SEQUENCE ID: NO: 1, or a polypeptide coded for by a polynucleotide sequence having 95% homology to SEQUENCE ID NO:1.
In yet still another embodiment, the present invention is directed to a method of screening for the ability of a compound to inhibit the production of inflammatory cytokine comprising determining the ability of the compound to bind to a polypeptide coded for by the polynucleotide sequence of SEQUENCE ID NO: 2, or a polypeptide coded for by a polynucleotide sequence having 95% homology to SEQUENCE ID NO: 2.
Especially preferred is the method wherein the inflammatory cytokine is interleukin 1.
In another embodiment, the present invention is directed to a method of treatment of a mammal having a disease condition characterized by an inflammatory component comprising administering to a mammal in need of such treatment a compound, or a pharmaceutically acceptable salt, ester, or prodrug of said compound, capable of binding to a polypeptide coded for by the polynucleotide sequence of SEQUENCE ID NO: 2, or a polypeptide coded for by a polynucleotide sequence having 95% homology to SEQUENCE ID NO: 2.
In yet still another embodiment, the present invention is directed to a pharmaceutical composition comprising a pharmaceutical carrier and a pharmaceutically active amount of a compound, or a pharmaceutically acceptable salt ester, or prodrug of said compound, said compound capable of binding to the polypeptide coded for by the polynucleotide sequence of SEQUENCE ID: NO: 2, or a polypeptide coded for by a polynucleotide sequence of having 95% homology to SEQUENCE ID NO:2.
In yet still another embodiment, the present invention is directed to a method of screening for the ability of a compound to inhibit the production of an inflammatory cytokine comprising determining the ability of the compound to bind to a polypeptide having the amino acid sequence of SEQUENCE ID NO: 3, or a polypeptide having 95% homology to SEQUENCE ID NO: 3.
Especially preferred is the method wherein the inflammatory cytokine is interleukin 1.
In another embodiment, the present invention is directed to a method of treatment of a mammal having a disease condition characterized by an inflammatory component comprising administering to a mammal in need of such treatment a compound, or a pharmaceutically acceptable salt, ester or prodrug of said compound, capable of binding to a polypeptide having the amino acid sequence of SEQUENCE ID NO: 3, or a polypeptide having 95% homology to SEQUENCE ID NO: 3.
In still another embodiment, the present invention is directed to a pharmaceutical composition comprising a pharmaceutical carrier and a pharmaceutically active amount of a compound, or pharmaceutically acceptable salt, ester, or prodrug of said compound, said compound capable of binding to the polypeptide of SEQUENCE ID: NO: 3, or a polypeptide having 95% homology to SEQUENCE ID NO: 3.
In yet still another embodiment, the present invention is directed to a method of screening for the ability of a compound to inhibit the production of an inflammatory cytokine comprising determining the ability of the compound to bind to a polypeptide having the amino acid sequence of SEQUENCE ID NO: 4, or a polypeptide having 95% homology to SEQUENCE ID NO: 4.
Especially preferred is the method wherein the inflammatory cytokine is interleukin 1.
In another embodiment, the present invention is directed to a method of treatment of a mammal having a disease condition characterized by an inflammatory component comprising administering to a mammal in need of such treatment a compound, or a pharmaceutically acceptable salt, ester, or prodrug of said compound, capable of binding to a polypeptide having the amino acid sequence of SEQUENCE ID NO: 4, or a polypeptide having 95% homology to SEQUENCE ID NO: 4.
In still another embodiment the present invention is directed to a pharmaceutical composition comprising a pharmaceutical carrier and a pharmaceutically active amount of a compound, or a pharmaceutically acceptable salt, ester, or prodrug of said compound, said compound capable of binding to the polypeptide having the amino acid sequence of SEQUENCE ID: NO: 4, or a polypeptide having 95% homology to SEQUENCE ID NO:4.
In another embodiment, the present invention is directed to the compound 1-(4-Chloro-2,6-diisopropyl-phenyl)-3-[2-fluoro-5-oxiranylbenzenesulfonyl]-urea.
In another embodiment, the present invention is directed to the compound 1-(1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)-3-[2-fluoro-5-oxiranylbenzenesulfonyl]-urea.